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Environmental & Socio-Economic Studies Environmental & Socio-economic Studies DOI: 10.1515/environ-2015-0046 Environ. Socio.-econ. Stud., 2014, 2, 4: 21-27 © 2014 Copyright by University of Silesia ________________________________________________________________________________________________ Micromorphological study (ultrastructure of lamina surface, seeds, ultrasculpture of pollen grains) of Gladiolus L. species (Iridaceae Juss.) of Ukrainian flora Svitlana L. Zhygalova1, Oksana A. Futorna1,2, Anatoliy Levanets3 1Department of Systematics and Floristics of Vascular Plants, M.G. Kholodny Institute of Botany, National Academy of Sciences of Ukraine, Tereschenkivska Str. 2, Kyiv 01601, Ukraine 2O.V. Fomin Botanical Garden, Educational-Scientific Centre “Institute of Biology”, National Taras Schevchenko University of Kyiv, S. Petlyury Str. 1, Kyiv 01032, Ukraine 3Research Unit for Environmental Sciences and Management, Botany Department, North-West University, Potchefstroom, Republic of South Africa, Research Unit for Environmental Sciences and Management, P.O. Box X6001, North-West University 2520, Republic of South Africa E–mail address: (corresponding author) [email protected] ________________________________________________________________________________________________________________________________________________ ABSTRACT Micro-morphological characteristics of the four Gladiolus L. species of the Ukrainian flora (G. imbricatus L., G. italicus Mill., G. palustris Gaudin and G. tenuis M. Bieb.) as regards leaves, seeds and pollens are presented with this investigation in a detailed way. An examination of the surface structure of the leaves, seeds and pollen grains of the Gladiolus species indicates that the characteristics of the ultrastructure of leaves and of pollen grains are not diagnostic for distinguishing species, but they could be important at genus level (leaves: features such as being amphistomatic, having the same quantity of immersed stomata on both surfaces and having a high stomata index, the presence and localisation of papillae, the shape of epidermal cells; pollen grains: monosulcate type with two operculums). However, the type of surface ultrastructure of the seed coat is a diagnostic feature as at genus level so for species. It can be mentioned that propose the use of features such as the shape and position of the cicatricle, the type of cuticle, the shape and boundaries of cells of testa, and the anticlinal cell walls as diagnostic features at genera level. The shape of seeds, the presence and disposition of wing, the level of the periclinal cell walls of the seed coat and types of relief are additional diagnostic features for distinguishing of Gladiolus species. KEY WORDS: Gladiolus, micromorphology, leaf lamina, seed, pollen ________________________________________________________________________________________________________________________________________________ 1. Introduction Micromorphological studies of the Iridaceae species lamina have been conducted by many The genus Gladiolus L. is the second largest authors (COLLINS, 1937; KARASAWA, 1942; SHORINA, genus of the family Iridaceae Juss after Iris L. 1975; RUDALL & MATHEW, 1990; RUDALL, 1991; According to the classification of GOLDBLATT & GOLDBLATT & LE THOMAS, 1997; EROL & KÜÇÜKER, MANNING (2008), the Gladiolus belongs to the 2003, 2007; KANDEMIR, 2009). However, the subfamily Crocoideae. The genus Gladiolus L. includes research data relating to the ultrastructure of the about 262 species that are found mainly in Africa leaves of the Ukrainian Gladiolus species are not and Madagascar, as well as in Europe, the Caucasus, available. central Asia, the Middle Asia and Asia Minor. Four The fruit and seed structure of Iridaceae was species are indicated for Ukrainian flora (MOSYAKIN studied by ARTYUSHENKO (1990) and RODIONENKO & FEDORONCHUK, 1999) – G. imbricatus L., G. italicus (1961). Later the seed micromorphology of 71 Mill., G. palustris Gaudin and G. tenuis M. Bieb. Iris species was investigated by Russian scientists Of these, Gladiolus imbricatus and G. tenuis are (RODIONENKO, 2005; ALEKSEEVA ET AL., 2007, 2011; noted as vulnerable in the RED DATA BOOK OF KRAVTSOVA & ZHYNKINA, 2008; ALEKSEEVA, 2010). UKRAINE (2009), and G. italicus and G. palustris are According to ALEKSEEVA ET AL. (2011), such considered endangered. researches are necessary because of the diagnostic 21 value of seed features at Iris species level. the Ukrainian flora and to investigate whether these MANNING & GOLDBLATT, (1991) studied the systematic characters are useful systematically. This is the first and phylogenetic significance of the seed testa of time such a study has been carried out in Ukraine. the African genera of Iridaceae – Nivenia Vent., Klattia Baker and Witsenia Thunb. EROL ET AL. 2. Materials and methods (2006) studied the seeds of nine Turkish Gladiolus species, including G. italicus. In Ukraine SZIKURA & The samples used for the study were SZIKURA (2003) detailed the characteristics of the herbarium specimens from the MG Kholodny fruit and seed testa of Iridaceae, including three Institute of Botany of NAS of Ukraine (KW). They Ukrainian Gladiolus species, but these results are were from a variety of habitats and were collected hardly reliable because of an incorrect selection in the wild. In order to study the ultrastructure of of samples (incorrect nomenclature and species the leaf lamina (middle third of leaf) and seeds, definition). the samples were mounted and sputter coated The literary data on the pollen morphology of with a thin layer of gold. To remove the wax from Iridaceae taxa is rather limited. RODIONENKO (1961) the leaf surface, each sample was placed in a xylol researched the pollen grains morphology of 73 solution for 24 hours. The study was carried out species of Iris, 24 species of Juno Tratt., five species using scanning electronic microscope JSM-6060 LA. of Crocus, one species of the genera Gynandriris The terminology used is, as far as possible, in Parl., Xiphium Mill. em. Rodion., Tigridia, accordance with works of ZAKHAREVICH (1954), Belamcanda, Sisyrinchium and five species of JUNIPER (1959), HALLAM (1970), HALLAM & CHAMBERS Gladiolus using light microscopy. The pollen grains (1970), CHUANG & HECKARD (1972), BARTHLOTT of Gladiolus, along with the size of the grains and (1981), JUNIPER & JEFFRY (1986), CHAKRABARTY & of the exine cells, are detailed by the author in a MUKHERJEE (1986), STERN (1992), BARTHLOTT ET AL. table. GOLDBLATT & LE THOMAS (1997) discussed the (1998). For the SEM study of the pollen grains’ phylogenetic reconstruction and classification of ultrasculpture (CEM, JSM 6060 LA), the samples the Afro-Madagascan genus Aristea after examining were mounted in 96% ethanol and then sputter the pollen grains using light and scanning electron coated with gold. The pollen grains were also microscopy. The ultrasculpture of pollen grains of examined under the light microscope (LM), 29 taxa of the genus Crocus were also studied by without acetolysis, and mounted in 30% glycerin Turkish scientists (IŞIK & DÖNMEZ, 2006; CANDAN (stained by fuchsine), but after rehydration of the ET AL., 2009; CANDAN & ÖZHATAY, 2013; and others). anthers in 50% glycerin. The pollen grains were These authors showed that the ultrasculpture of described using the terminology of PUNT ET AL. the pollen grains of the Turkish Crocus species is (1994) and TOKAREV (2002). similar to Europe’s Crocus species. The pollen morphology of 10 Iris species from the Far East 2.1. List of samples was examined by BOLTENKOV & GRIGORJEVA (2012) using a light and scanning electron microscope. G. іmbricatus L.: 1. Chernivtsi Region, Ghertsaiv However, the data on the pollen morphology District, between villages Kulykivka and Tarnavka, of the Gladiolus species is limited. BEUG (1963) meadow. 03.07.2005. A. Tokaryuk, O. Volutsa (KW), described the pollen grains of five Gladiolus 2. Ivano-Frankivsk Region, Nadvirnyansk District, species and, as mentioned above, RODIONENKO east border of village Mykulychin, meadow, 1059 m also studied the pollen grains of five species. a.s.l. 24.08.2010. L. Borsukevich, K. Danylyuk (KW), Furthermore, BOBROV ET AL. (1983) characterised 3. Chernihiv Region, Oster District, environs of the pollen grains of G. imbricatus, while the village Yevminka, meadow along the Desna River. palynomorphological data of Gladiolus is cited in 24.06.2007. O. Bulakh, S. Zhygalova (KW), 4. Sumy the monograph “The Iris family” (GOLDBLATT & Region, Putivl District, beam near village Shyryajevo. MANNING, 2008). However, such research is 21.06.2009. Ya. Didukh (KW), 5. Transcarpathian absent in Ukraine. We have already studied the Region, urban village Jasinya, wet meadow at the micromorphological characteristics of G. imbricatus hill. 19.07.2008. I. Olshanskij, S. Zhygalova (KW). of Ukrainian flora (ZHYGALOVA & FUTORNA, 2013) and our investigation indicated the necessity of G. tenuis M. Bieb.: 1. Poltava Region, Kobelyaky complex micromorphological study of all Ukrainian District, environs of the biostation “Luchki” of the Gladiolus species. Poltava Pedagogical Institute, reserve tract The aim of this study is to give a complete “Sokilske”, meadow after logged land. 18.06.1994. description of the surface structure of leaves, N.O. Stetsyuk (KW), 2. Chernigiv Region, Sosnytsya seeds and pollen grains of the Gladiolus species of District, 1 km on the west from the village 22 Yaklychi, meadow (with dominating Festuca G. palustris Gaudin.: Ivano-Frankivsk Region, pratensis and Deschampsia cespitosa)
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